This Mouse CD200R overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD200R protein (Cat: 50209-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Pro 238) of mouse CD200R1 (NP_067300.1) was expressed, with a C-terminal polyhistidine tag.
The recombinant mouse CD200R1 consists of 224 amino acids after removal of the signal peptide and has a predicted molecular mass of 24.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rmCD200R1 is approximately 55-65 kDa due to glycosylation.
Mouse CD200R HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
The cluster of differentiation (CD) system is commonly used as cell markers in Immunophenotyping. Different kinds of cells in the immune system can be identified through the surface CD molecules associating with the immune function of the cell. There are more than 320 CD unique clusters and subclusters have been identified. Some of the CD molecules serve as receptors or ligands important to the cell through initiating a signal cascade which then alter the behavior of the cell. Some CD proteins do not take part in cell signal process but have other functions such as cell adhesion. Cell surface glycoprotein CD200 receptor 1 (CD200R1) is an isoform of CD200 receptors that is expressed on cells of the myeloid lineage. CD200R1 is a receptor for the OX-2 membrane glycoprotein. The receptor-substrate interaction may serve as a myeloid downregulatory signal.